Autologous HIV-1 resistant T cells through accelerated CCR5 gene disruption

通过加速 CCR5 基因破坏产生自体 HIV-1 抗性 T 细胞

• 批准号: 8603742
• 负责人: Alexander Astrakhan
• 金额: $ 50.77万
• 依托单位: PRECISION GENOME ENGINEERING, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8603742
• 关键词: Adverse effects; Alleles; Anti-Retroviral Agents; Autologous; Biodistribution; Biological; Biological Assay; Bone Marrow Transplantation; CCR5 gene; CD4 Positive T Lymphocytes; Case Study; Cell Separation; Cells; Characteristics; Clinic; Clinical; Clinical Trials; DNA; Data Set; Dependence; Development; Dose; Electroporation; Engineering; Engraftment; Enhancement Technology; Evaluation; Event; Exonuclease; Gene Delivery; Gene Targeting; Gene-Modified; Generations; Genes; Genetic; Genome; Genomics; HIV; HIV Infections; HIV-1; Health; Heterogeneity; Highly Active Antiretroviral Therapy; Homing; Human; Immunodeficient Mouse; In Vitro; Individual; Infection; Lead; Length; Lymphocyte; Mediating; Messenger RNA; Methods; Modeling; Monitor; Mutation; Patients; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Positioning Attribute; Process; Production; Protocols documentation; Qualifying; Quality of life; Reagent; Regimen; Resistance; Safety; Site; Small Business Technology Transfer Research; Specificity; Staging; Surface; T cell therapy; T-Lymphocyte; Technology; Testing; Therapeutic; Toxic effect; Toxicology; Transcription Coactivator; Validation; Variant; Viral; Viremia; alternative treatment; base; endonuclease; improved; in vivo; manufacturing process; mouse model; novel; nuclease; outcome forecast; peripheral blood; potency testing; pre-clinical; preclinical efficacy; preclinical safety; product development; programs; public health relevance; receptor; reconstitution; research study; success

DESCRIPTION (provided by applicant): Treating patients infected with Human Immunodeficiency Virus-1 (HIV-1) is a substantial clinical challenge with evolving pharmaceutical drug and dosing options. Treatments should improve the health and quality of life for HIV-infected individuals by reducing viremia and preserving immunological function. Currents therapeutic approaches utilize combination antiretroviral drug regiments (cARV, historically referred to highly active antiretroviral therapy, or HAART) to reduce ongoing viral replication and improve the patient's prognosis. However, cARV therapies are not curative and furthermore they are expensive, are associated with high rates of drug-related toxicity, and are highly susceptible to viral evasion through resistance-conferring mutation. Genetic targeting CCR5 is a promising alternative approach to cARV therapy. The CCR5 gene is used by HIV-1 for cellular entry and those individuals who naturally lack CCR5 expression are fully resistant to HIV-1 infection. In an important clinical case study, bone marrow transplantation from a CCR5-deficient donor has produced the first recorded case of functional cure of an ongoing HIV-1 infection. This has prompted therapeutic strategies to create HIV-1 resistant patient-derived T cells using nuclease- based reagents to create disruptive mutations in the CCR5 gene. Though providing critical proof-of- concept in HIV-1 patients, the success of the gene-targeting therapy currently in clinical trials appears to be limited due to low gene disruption activity of these firt-generation nuclease technologies. Pregenen has used its surface display engineering platform to produce highly specific CCR5- targeting LAGLIDADG homing endonucleases (LHE), recently reformatted as extremely powerful MegaTAL nucleases and combined with an additional layer of rate-enhancement technology - exonuclease-accelerated genome editing (X-AGE) developed at SCRI. The proposed therapy will isolated and process patient-derived T-cells by delivering these enhanced reagents using a simplified and scalable mRNA-based method. This manufacturing process will be used to create HIV-resistant, CCR5-deficient autologous CD4+ T cell. Autologous CCR5-disrupted T cells have the potential to durably suppress HIV-1 infection and reconstitute the full functionality of the CD4+ compartment while reducing dependence on antiretroviral drugs. The experiments described in this proposal are part of a comprehensive plan to bring the CCR5-targeting nuclease therapy to the clinic. We plan to profile the safety, biodistribution, and potency of primary human T cells treated with CCR5-targeting reagents. The best-performing reagents and protocols will be used to generate CCR5-deficient human CD4+ T cells to test the capacity of these cells to withstand HIV infection in vitro and in vivo. We will se this data-set to support pivotal preclinical experiments and subsequent IND submission for the continued clinical development of our cellular therapy.

描述（由申请人提供）：治疗感染人类免疫缺陷病毒-1（HIV-1）的患者是一项重大的临床挑战，药物和剂量选择不断发展。治疗应通过减少病毒血症和保护免疫功能来改善艾滋病毒感染者的健康和生活质量。目前的治疗方法利用组合抗逆转录病毒药物方案（cARV，历史上称为高效抗逆转录病毒疗法，或HAART）来减少正在进行的病毒复制并改善患者的预后。然而，cARV疗法不是治愈性的，而且它们是昂贵的，与高比率的药物相关毒性相关，并且通过赋予抗性的突变对病毒逃避高度敏感。基因靶向CCR 5是cARV治疗的一种有前途的替代方法。CCR 5基因被HIV-1用于细胞进入，那些天然缺乏CCR 5表达的个体完全抵抗HIV-1感染。在一项重要的临床病例研究中，来自CCR 5缺陷供体的骨髓移植产生了第一例持续HIV-1感染的功能性治愈病例。这促使了使用基于核酸酶的试剂在CCR 5基因中产生破坏性突变来产生HIV-1抗性患者来源的T细胞的治疗策略。虽然在HIV-1患者中提供了关键的概念验证，但由于这些第一代核酸酶技术的低基因破坏活性，目前在临床试验中的基因靶向治疗的成功似乎是有限的。Pregenen使用其表面展示工程平台生产高度特异性的CCR 5靶向LAGLIDADG归巢核酸内切酶（LHE），最近重新格式化为非常强大的MegaTAL核酸酶，并结合了额外的一层速率增强技术-SCRI开发的核酸外切酶加速基因组编辑（X-AGE）。所提出的疗法将通过使用简化和可扩展的基于mRNA的方法递送这些增强的试剂来分离和处理患者来源的T细胞。该制造工艺将用于制造抗艾滋病毒、缺乏CCR 5的自体CD 4 + T细胞。自体CCR 5破坏的T细胞有可能持久地抑制HIV-1感染，并重建CD 4+区室的全部功能，同时减少对抗逆转录病毒药物的依赖。本提案中描述的实验是将CCR 5靶向核酸酶疗法引入临床的综合计划的一部分。我们计划分析用CCR 5靶向试剂处理的原代人T细胞的安全性、生物分布和效力。性能最好的试剂和方案将用于产生CCR 5缺陷的人CD 4 + T细胞，以测试这些细胞在体外和体内抵抗HIV感染的能力。我们将使用该数据集来支持关键的临床前实验和随后的IND提交，以继续我们的细胞疗法的临床开发。